目的 对济南市两株输入性登革热病例进行血清学和基因检测,获取全基因数据并进行分子特征分析.方法 采集济南市2例登革热疑似阳性病例的全血样本,通过Real-time PCR对病毒进行分型检测.离心分离血清,用于胶体金法检测登革病毒NS1抗原、IgM抗体和IgG抗体;同时扩增病毒全基因序列,将序列与国内外分离株序列进行比对,Mega 7.0软件构建进化树,RDP4软件进行基因重组分析.结果 2例病例核酸分型检测结果为DENV3型,胶体金检测结果显示NS1抗原均为阳性,IgG抗体结果均为阴性;测序后拼接序列长度分别为10 707 nt和10 706 nt,两条序列仅在末端有6个核苷酸的差异,经系统进化分析,这两例病毒株属于DENV3型基因1型,与2017年孟加拉国流行株具有高度同源性.结论 济南市登革热的防控依旧需要加强输入性病例的监测.
This study was aimed at identifying the serotypes and genotypes of two dengue fever virus strains isolated from two imported cases of dengue fever in Jinan city;obtaining their whole genome sequences;and analyzing their molecular evolu-tionary characteristics.Whole blood samples from two suspected positive cases of dengue fever in Jinan city were collected,and the subtypes were identified by real-time PCR.Serum samples were collected to detect dengue virus IgM and IgG antibodies,and NS1 antigen,though the colloidal gold method.Viral whole genome fragments were amplified and sequenced,and com-pared against reference sequences of viruses isolated in China and other countries.Phylogenetic trees were constructed in Mega 7.0 software,and gene recombination analysis was performed in RDP4 software.The viruses isolated from whole blood samples were identified as dengue virus type Ⅲ.Colloidal gold tests indicated that the suspected dengue fever cases were positive for NS1 and negative for IgG.The two complete genomes were sequenced and found to be 10 707 nt and 10 706 nt.The two se-quences differed in only 6 nucleotides.On the basis of phylogenetic analysis,these two virus strains were grouped into Dengue virus type 3 genotype 1,and were found to have high homology and close relationships with the 2017 Bangladesh genome.Sur-veillance of imported cases should be strengthened to prevent and control dengue fever epidemics.
韩莹;焦海涛;刘子薇;武晶晶;刘岚铮
济南市疾病预防控制中心,济南 250021
基础医学
登革病毒;全基因序列;系统进化分析
dengue fever virus;whole genome sequence;phylogenetic analysis
《中国人兽共患病学报》 2024 (003)
253-256 / 4
10.3969/j.issn.1002-2694.2024.00.036
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